Regulation of the Innate Immune Response by Fibronectin: Synergism between the III-1 and EDA Domains
Figure 6
IL-8 expression induced by FnIII domains is NF-κB and TLR4-dependent.
Fibroblasts were serum-starved overnight before treatment with 20 µM FnIII-1c, FnEDA, FnIII-10n, FnIII-13 or PBS in 0.1% BSA-DMEM for the indicated amount of time (A) or for 4 h at the indicated dose (B). Fibroblasts or THP-1 monocytes were incubated with increasing doses of LPS for 4 h (C). Tenascin C (100 µg/ml), LPS (1 µg/ml) or FnEDA (200 µg/ml) were incubated with either dermal fibroblasts or THP-1 monocytes for 4 h (D). Cells were pretreated with the indicated amounts of inhibitor or monoclonal antibody for 1 h prior to the addition of FnIII-1c (E) or FnEDA (F) in 0.1% BSA-DMEM for 4 h. In all experiments, conditioned media was collected and IL-8 protein amounts determined by ELISA. To determine significance, statistical analysis was performed using Student's t test. *p<0.05. Error bars indicate mean ± S.D. of three separate experiments performed in triplicate.